Harnessing the endogenous Type I-C CRISPR-Cas system for genome editing in Bifidobacterium breve

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dc.check.date06/08/2024en
dc.check.infoAccess to this article is restricted until 6 months after publication by request of the publisheren
dc.contributor.authorHan, Xiaoen
dc.contributor.authorChang, Luluen
dc.contributor.authorChen, Haiqinen
dc.contributor.authorZhao, Jianxinen
dc.contributor.authorTian, Fengweien
dc.contributor.authorRoss, R. Paulen
dc.contributor.authorStanton, Catherineen
dc.contributor.authorvan Sinderen, Douween
dc.contributor.authorChen, Weien
dc.contributor.authorYang, Boen
dc.contributor.editorDudley, Edward G.en
dc.contributor.funderNational Key Research and Development Program of Chinaen
dc.contributor.funderNational Natural Science Foundation of Chinaen
dc.contributor.funderProject 211en
dc.contributor.funderCollaborative Innovationcenter of Food Safety and Quality Control in Jiangsu Provinceen
dc.date.accessioned2024-04-18T14:24:51Z
dc.date.available2024-04-18T14:24:51Z
dc.date.issued2024-02-06en
dc.description.abstractBifidobacterium breve, one of the main bifidobacterial species colonizing the human gastrointestinal tract in early life, has received extensive attention for its purported beneficial effects on human health. However, exploration of the mode of action of such beneficial effects exerted by B. breve is cumbersome due to the lack of effective genetic tools, which limits its synthetic biology application. The widespread presence of CRISPR-Cas systems in the B. breve genome makes endogenous CRISPR-based gene editing toolkits a promising tool. This study revealed that Type I-C CRISPR-Cas systems in B. breve can be divided into two groups based on the amino acid sequences encoded by cas gene clusters. Deletion of the gene coding uracil phosphoribosyl-transferase (upp) was achieved in five B. breve strains from both groups using this system. In addition, translational termination of uracil phosphoribosyl-transferase was successfully achieved in B. breve FJSWX38M7 by single-base substitution of the upp gene and insertion of three stop codons. The gene encoding linoleic acid isomerase (bbi) in B. breve, being a characteristic trait, was deleted after plasmid curing, which rendered it unable to convert linoleic acid into conjugated linoleic acid, demonstrating the feasibility of successive editing. This study expands the toolkit for gene manipulation in B. breve and provides a new approach toward functional genome editing and analysis of B. breve strains.en
dc.description.sponsorshipNational Key Research and Development Program of China (2022YFA0912200); National Natural Science Foundation of China (32021005); 111 Project (BP0719028)en
dc.description.statusPeer revieweden
dc.description.versionPublished Versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.articleide02074-23en
dc.identifier.citationHan, X., Chang, L., Chen, H., Zhao, J., Tian, F., Ross, R. P., Stanton, C., van Sinderen, D., Chen, W. and Yang, B. (2024) 'Harnessing the endogenous Type I-C CRISPR-Cas system for genome editing in Bifidobacterium breve', Applied and Environmental Microbiology, 90(3), e02074-23 (16pp). https://doi.org/10.1128/aem.02074-23en
dc.identifier.doihttps://doi.org/10.1128/aem.02074-23en
dc.identifier.eissn1098-5336en
dc.identifier.endpage16en
dc.identifier.issn0099-2240en
dc.identifier.issued3en
dc.identifier.startpage1en
dc.identifier.urihttps://hdl.handle.net/10468/15813
dc.identifier.volume90en
dc.language.isoenen
dc.publisherAmerican Society for Microbiologyen
dc.rights© 2024, American Society for Microbiology. All Rights Reserved.en
dc.subjectBifidobacterium breveen
dc.subjectCRISPR-Casen
dc.subjectType I-Cen
dc.subjectGenome editingen
dc.titleHarnessing the endogenous Type I-C CRISPR-Cas system for genome editing in Bifidobacterium breveen
dc.typeArticle (peer-reviewed)en
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